Science for Life Laboratory, School of Engineering Sciences in Chemistry, Biotechnology and Health, KTH - Royal Institute of Technology, 17121 Stockholm, Sweden.
Department of Immunology, Genetics and Pathology, Rudbeck Laboratory, Uppsala University, 75185 Uppsala, Sweden.
J Proteome Res. 2020 Dec 4;19(12):4766-4781. doi: 10.1021/acs.jproteome.0c00486. Epub 2020 Nov 10.
The localization of proteins at a tissue- or cell-type-specific level is tightly linked to the protein function. To better understand each protein's role in cellular systems, spatial information constitutes an important complement to quantitative data. The standard methods for determining the spatial distribution of proteins in single cells of complex tissue samples make use of antibodies. For a stringent analysis of the human proteome, we used orthogonal methods and independent antibodies to validate 5981 antibodies that show the expression of 3775 human proteins across all major human tissues. This enhanced validation uncovered 56 proteins corresponding to the group of "missing proteins" and 171 proteins of unknown function. The presented strategy will facilitate further discussions around criteria for evidence of protein existence based on immunohistochemistry and serves as a useful guide to identify candidate proteins for integrative studies with quantitative proteomics methods.
蛋白质在组织或细胞类型特异性水平上的定位与蛋白质功能密切相关。为了更好地了解每个蛋白质在细胞系统中的作用，空间信息是定量数据的重要补充。确定复杂组织样本中单细胞中蛋白质空间分布的标准方法是利用抗体。为了严格分析人类蛋白质组，我们使用正交方法和独立抗体来验证了 5981 种抗体，这些抗体在所有主要人类组织中均显示出 3775 种人类蛋白质的表达。这种增强的验证揭示了 56 种对应于“缺失蛋白质”组的蛋白质和 171 种未知功能的蛋白质。所提出的策略将有助于进一步围绕基于免疫组织化学的蛋白质存在证据的标准展开讨论，并为使用定量蛋白质组学方法进行综合研究确定候选蛋白质提供有用的指导。
